Journal
NUCLEIC ACIDS RESEARCH
Volume 48, Issue 3, Pages -Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkz1141
Keywords
-
Categories
Funding
- German Science Foundation [CRC992, CRC1381, 403222702]
- Germany's Excellence Strategy (CIBSS -EXC-2189) [390939984]
- Max Planck Research Group Leader Program
Ask authors/readers for more resources
Determination of the in vivo binding sites of RNA-binding proteins (RBPs) is paramount to understanding their function and how they affect different aspects of gene regulation. With hundreds of RNA-binding proteins identified in human cells, a flexible, high-resolution, high-throughput, highly multiplexible and radioactivity-free method to determine their binding sites has not been described to date. Here we report FLASH (Fast Ligation of RNA after some sort of Affinity Purification for High-throughput Sequencing), which uses a special adapter design and an optimized protocol to determine protein-RNA interactions in living cells. The entire FLASH protocol, starting from cells on plates to a sequencing library, takes 1.5 days. We demonstrate the flexibility, speed and versatility of FLASH by using it to determine RNA targets of both tagged and endogenously expressed proteins under diverse conditions in vivo.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available